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Article Open Access

PTPN18 functions as a tumor suppressor in breast cancer by negatively regulating cyclin E

  • Authors:
    • Na Zhang
    • Tao Wang
    • Bin Bai
    • Xiaonan Zhang
    • Wenying Xu
    • Weilu Chen
    • Yang Yu
    • Bing Wang
  • View Affiliations / Copyright

    Affiliations: Institute of Biochemistry and Molecular Biology, College of Life and Health Sciences, Northeastern University, Shenyang, Liaoning 110169, P.R. China
    Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 30
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    Published online on: December 30, 2025
       https://doi.org/10.3892/ijo.2025.5843
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Abstract

Protein tyrosine phosphatase non‑receptor 18 (PTPN18) is widely expressed in breast cancer (BC) cell lines. Additionally, high levels of PTPN18 facilitate an improved overall survival and prognosis in patients with BC. However, the effects and mechanisms of PTPN18 in BC remain unclear. In the present study, it was found that PTPN18 serves a tumor suppressor role in BC cells by promoting apoptosis, inhibiting proliferation and metastasis and inducing cell cycle arrest. Bioinformatics analysis showed that PTPN18 was significantly negatively correlated with the cell cycle and downregulated cyclin E expression, which was consistent with the experimental results. Subsequent co‑immunoprecipitation assay results showed that PTPN18 could bind to cyclin E and promote its degradation through the ubiquitin‑proteasome pathway. Moreover, the addition of cyclin E2 did not reduce the binding of PTPN18 to cyclin E1. In the present study, the signaling pathways involved in cell cycle regulation were further investigated and it was found that PTPN18 may regulate the expression levels of cyclin‑dependent kinase (CDK) inhibitor 1A and CDK inhibitor 1B proteins through phosphatidylinositol 3‑kinase/protein kinase B signaling pathway, which leads to cell cycle arrest and tumor inhibition in BC. Thus, analysis of the tumor suppressor mechanism of PTPN18 not only helps us to understand its biological function but also provides a theoretical basis for the development of new therapeutic strategies for BC.
View Figures

Figure 1

High expression of PTPN18 improves
the survival and prognosis of patients with BC. (A) The UALCAN
database was used to analyze the protein expression changes of
PTPN18 in BC. (B) Box plots of PTPN18 expression in different tumor
stages using data from the GEPIA database. (C) Kaplan-Meier plotter
was used to analyze the effect of high and low PTPN18 expression on
the OS of patients with BC. (D) Effect of PTPN18 expression on the
relapse-free survival of patients with BC. (E) GEPIA database was
used to analyze the effect of high and low PTPN18 expression on the
OS of patients with BC. (F) GEPIA database was used to analyze the
influence of PTPN18 expression on disease-free survival in patients
with BC. BC, breast cancer; PTPN18, protein tyrosine phosphatase
non-receptor 18; GEPIA, Gene Expression Profiling Interactive
Analysis. OS, overall survival.

Figure 2

PTPN18 suppresses the invasion and
migration of BC cells. (A) Representative images of the Transwell
invasion assays of BC cells with PTPN18 overexpression or
knockdown. Cells were collected after 48 h of culture. Scale bars,
200 µm. (B) Statistical analysis of the number of invasive
cells. (C) Cell migration capability was determined with a wound
healing assay. Scale bars, 200 µm. (D) Statistical analysis
of the wound area. (E) Expression of genes associated with cell
migration and invasion measured by reverse
transcription-quantitative polymerase chain reaction. (F)
Immunoblot analysis of the epithelial-mesenchymal transition
marker, E-cadherin, following interference with PTPN18 expression.
RNA or protein was extracted 48 h after overexpression and 72 h
after knockdown transfections. Data are shown as the mean ± SD from
three technical replicates. *P<0.05,
**P<0.01 and ***P<0.001. PTPN18,
protein tyrosine phosphatase non-receptor 18; BC, breast cancer;
ns, not significant; siRNA, small interfering RNA.

Figure 3

PTPN18 inhibits cell proliferation
and induces apoptosis in breast cancer cells. (A) Cell viability
analysis at different time points following PTPN18 overexpression.
(B) Assessment of cell viability over time upon PTPN18 knockdown.
(C) Representative images from the colony formation assays after 2
weeks of cultivation. (D) Statistical analysis of the number of
colonies counted using ImageJ. (E) Apoptosis scatter plots based on
PI and Annexin V-FITC staining were obtained by flow cytometry. (F)
Quantitative analysis of apoptosis rate measured by flow cytometry.
(G) DAPI was used to stain the nuclei. The nuclei of apoptotic
cells showed dense hyperchromatic bright spots under a fluorescence
microscope. Scale bars, 100 µm. (H) Reverse
transcription-quantitative polymerase chain reaction analysis of
the mTOR, Bcl-2 and Bax mRNA expression levels. (I) Protein
expression levels of mTOR, Bcl-2 and Bax were analyzed by western
blotting following interference with PTPN18 expression. Data are
shown as the mean ± SD from three technical replicates following
overexpression (48 h) or knockdown (72 h) treatment.
*P<0.05, **P<0.01 and
***P<0.001. PTPN18, protein tyrosine phosphatase
non-receptor 18; mTOR, mechanistic target of rapamycin kinase; ns,
not significant; siRNA, small interfering RNA.

Figure 4

PTPN18 induces S phase cell cycle
arrest in breast cancer cells by downregulating cyclin E
expression. (A and B) DNA content was measured by flow cytometry to
analyze cell cycle distribution under PTPN18 (A) overexpression and
(B) knockdown conditions. (C and D) Statistical analysis percentage
of cells at different cell cycle stages in (C) Panel A and (D)
Panel B. (E) Reverse transcription-quantitative polymerase chain
reaction was used to analyze the effect of PTPN18 on cyclin mRNA
expression levels. (F) Impact of PTPN18 on the transcriptional
levels of CDKs. (G) Effect of PTPN18 on cyclin and CDK protein
expression levels. (H) Statistics of changes in cell cycle-related
protein expression. The bar graph was obtained by normalizing the
expression levels to β-actin. All data were acquired 48 h after
overexpression or 72 h after knockdown and are shown as the mean ±
SD from three technical replicates. **P<0.01 and
***P<0.001. PTPN18, protein tyrosine phosphatase
non-receptor 18; CDKs, cyclin-dependent kinases; ns, not
significant.

Figure 5

Bioinformatic statistical analysis
showed a significant negative correlation between PTPN18 and the
cell cycle in BC. (A) Bar chart of the standardized enrichment
score of PTPN18 in BC. (B) Cell cycle gene set enrichment analysis
of PTPN18 in BC. (C) Volcano plot showing the genes associated with
PTPN18 in BC samples analyzed by LinkedOmics. (D) Correlation
analysis of PTPN18 with cyclins E1 and E2 using data from the Gene
Expression Profiling Interactive Analysis database. (E) Expression
profiles of the cyclin E1 and cyclin E2 genes in normal and tumor
tissues of the breast (T, red; N, gray). (F) Box plots of gene
expression changes in different tumor stages. (G and H)
Kaplan-Meier Plotter database was used to analyze the effect of the
cyclin E1 or cyclin E2 expression levels on the (G) overall
survival and (H) relapse-free survival of patients with BC. PTPN18,
protein tyrosine phosphatase non-receptor 18; BC, breast cancer; T,
tumor; N, normal.

Figure 6

PTPN18 can interact with cyclin E1
and promote its proteasomal degradation. (A) Effect of PTPN18 on
the cyclin E1 protein levels at different time points of CHX (10
µg/ml) treatment. (B) Knockdown of PTPN18 suppressed cyclin
E1 degradation under CHX-treated conditions. (C) Proteasome
inhibitor MG132 (10 µM) significantly decreased the effects
of PTPN18 on cyclin E1 degradation. (D) Effect of PTPN18 knockdown
on cyclin E1 expression in the absence or presence of MG132
treatment. The bar graph was obtained by normalizing the levels to
β-actin. (E and F) Combined treatment with CHX and MG132 almost
abolished the effects of PTPN18 (E) overexpression and (F)
knockdown on cyclin E1 protein levels compared with controls. (G)
PTPN18 overexpression elevated the ubiquitination level of cyclin
E1. (H) PTPN18 knockdown decreased cyclin E1 ubiquitination. (I)
Ubiquitination of cyclin E1 by PTPN18 requires enzymatic activity.
(J) Cyclin E1 is dephosphorylated by PTPN18 upon EGF (100 ng/ml, 30
min) stimulation. (K) PTPN18 knockdown upregulated the tyrosine
phosphorylation levels of cyclin E1. (L and M) Analysis of the
results from (L) forward and (M) reverse co-immunoprecipitation
showed the interaction between ectopic PTPN18 and cyclin E1. (N)
The physical association between endogenous PTPN18 and cyclin E1
was validated using co-immunoprecipitation. Data are shown as the
mean ± SD values from three technical replicates following 48 h of
overexpression or 72 h of knockdown treatment.
*P<0.05, **P<0.01 and
***P<0.001. PTPN18, protein tyrosine phosphatase
non-receptor 18; CHX, cycloheximide; ns, not significant.

Figure 7

PTPN18 promotes the ubiquitinated
degradation of cyclin E2. (A) Effect of PTPN18 on cyclin E2 protein
expression at different time points of CHX (10 µg/ml)
treatment. (B) Knockdown of PTPN18 inhibits the degradation of
cyclin E2. (C) Effect of PTPN18 overexpression on cyclin E2
expression in the absence or presence of MG132 (10 µM, 6 h)
treatment. (D) Effect of PTPN18 knockdown on cyclin E2 expression
in the absence or presence of MG132 (10 µM, 6 h) treatment.
(E) Overexpression or (F) knockdown of PTPN18 had no significant
effect on the cyclin E2 protein levels compared with the controls
when CHX and MG132 were combined. (G) Overexpression of PTPN18
enhanced cyclin E2 ubiquitination. (H) PTPN18 knockdown decreased
the ubiquitination level of cyclin E2. (I) PTPN18 CS reduced the
ubiquitination of cyclin E2. (J) PTPN18 regulated the pan-tyrosine
phosphorylation levels of cyclin E2. (K) PTPN18 knockdown increased
the tyrosine phosphorylation levels of cyclin E2. The binding
between exogenous PTPN18 and cyclin E2 was verified by
co-immunoprecipitation: (L) immunoprecipitation with PTPN18-HA and
(M) immunoprecipitation with flag-cyclin E2. (N) Cyclin E2 had no
significant effect on the binding of PTPN18 to cyclin E1. Data are
shown as the mean ± SD from three technical replicates following 48
h of overexpression or 72 h of knockdown transfection.
***P<0.001. ns, not significant; PTPN18, protein
tyrosine phosphatase non-receptor 18; CHX, cycloheximide; CS,
C229S; ns, not significant; si-, small interfering.

Figure 8

PTPN18 can regulate the expression
levels of CDKN1A and CDKN1B proteins through the PI3K/AKT signaling
pathway leading to cell cycle arrest. (A and B) STRING database
analysis showed proteins that interact with (A) cyclin E1 (CCNE1)
and (B) E2 (CCNE2). (C) GEPIA database analysis of the CDKN1A and
CDKN1B association with cyclin E1. (D) Correlation analysis between
CDKN1A and cyclin E2 using the GEPIA database. (E) Western blot
analysis of the PI3K/AKT signaling pathway protein expression
levels in MCF-7 cells. Levels of P-PI3K p85, P-AKT and P-GSK3 were
monitored after a 30-min stimulation with EGF (100 ng/ml). (F)
Relative quantitative analysis of the PI3K/AKT signaling pathway
protein expression levels. (G) Western blotting demonstrated the
expression levels of FOXO1, PI3K p110β, CDKN1A and CDKN1B. (H) The
bar graph was obtained by normalizing the levels to β-actin. Data
are shown as the mean ± SD from three technical replicates
following 48 h of overexpression or 72 h of knockdown.
**P<0.01 and ***P<0.001. PTPN18,
protein tyrosine phosphatase non-receptor 18; GEPIA, Gene
Expression Profiling Interactive Analysis; PI3K,
phosphatidylinositol 3-kinase; AKT, protein kinase B; CDKN1A
(p21Cip1), cyclin-dependent kinase inhibitor 1A; CDKN1B (p27Kip1),
cyclin-dependent kinase inhibitor 1B; P-, phosphorylation; FOXO1,
forkhead box O1; ns, not significant; P-, phosphorylated; siRNA,
small interfering RNA.

Figure 9

Proposed working model in which
PTPN18 negatively regulates cyclin E to interfere with cell cycle
progression to exert tumor suppressive effects in BC. The present
study found that PTPN18 could inhibit the AKT signaling pathway by
downregulating the expression of PI3K p110β as well as the tyrosine
phosphorylation of PI3K p85 and AKT. The inhibited AKT pathway
weakens the negative regulation of downstream FOXO1, CDKN1A and
CDKN1B. Increased expression of CDKN1A and CDKN1B enhances the
inhibition of the cell cycle and cyclin E expression, ultimately
leading to cell cycle arrest and BC suppression. PTPN18 can also
enhance cell cycle arrest by downregulating cyclin E expression
through the ubiquitin-proteasome pathway. PTPN18, protein tyrosine
phosphatase non-receptor 18; BC, breast cancer; PI3K,
phosphatidylinositol 3-kinase; AKT, protein kinase B; CDKN1A
(p21Cip1), cyclin-dependent kinase inhibitor 1A; CDKN1B (p27Kip1),
cyclin-dependent kinase inhibitor 1B; FOXO1, forkhead box O1.
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Copy and paste a formatted citation
Spandidos Publications style
Zhang N, Wang T, Bai B, Zhang X, Xu W, Chen W, Yu Y and Wang B: PTPN18 functions as a tumor suppressor in breast cancer by negatively regulating cyclin E. Int J Oncol 68: 30, 2026.
APA
Zhang, N., Wang, T., Bai, B., Zhang, X., Xu, W., Chen, W. ... Wang, B. (2026). PTPN18 functions as a tumor suppressor in breast cancer by negatively regulating cyclin E. International Journal of Oncology, 68, 30. https://doi.org/10.3892/ijo.2025.5843
MLA
Zhang, N., Wang, T., Bai, B., Zhang, X., Xu, W., Chen, W., Yu, Y., Wang, B."PTPN18 functions as a tumor suppressor in breast cancer by negatively regulating cyclin E". International Journal of Oncology 68.3 (2026): 30.
Chicago
Zhang, N., Wang, T., Bai, B., Zhang, X., Xu, W., Chen, W., Yu, Y., Wang, B."PTPN18 functions as a tumor suppressor in breast cancer by negatively regulating cyclin E". International Journal of Oncology 68, no. 3 (2026): 30. https://doi.org/10.3892/ijo.2025.5843
Copy and paste a formatted citation
x
Spandidos Publications style
Zhang N, Wang T, Bai B, Zhang X, Xu W, Chen W, Yu Y and Wang B: PTPN18 functions as a tumor suppressor in breast cancer by negatively regulating cyclin E. Int J Oncol 68: 30, 2026.
APA
Zhang, N., Wang, T., Bai, B., Zhang, X., Xu, W., Chen, W. ... Wang, B. (2026). PTPN18 functions as a tumor suppressor in breast cancer by negatively regulating cyclin E. International Journal of Oncology, 68, 30. https://doi.org/10.3892/ijo.2025.5843
MLA
Zhang, N., Wang, T., Bai, B., Zhang, X., Xu, W., Chen, W., Yu, Y., Wang, B."PTPN18 functions as a tumor suppressor in breast cancer by negatively regulating cyclin E". International Journal of Oncology 68.3 (2026): 30.
Chicago
Zhang, N., Wang, T., Bai, B., Zhang, X., Xu, W., Chen, W., Yu, Y., Wang, B."PTPN18 functions as a tumor suppressor in breast cancer by negatively regulating cyclin E". International Journal of Oncology 68, no. 3 (2026): 30. https://doi.org/10.3892/ijo.2025.5843
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